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Genotypic and phenotypic modifications of Neisseria meningitidis after an accidental human passage.

Omer H, Rose G, Jolley KA, Frapy E, Zahar JR, Maiden MC, Bentley SD, Tinsley CR, Nassif X, Bille E - PLoS ONE (2011)

Bottom Line: By comparing the number of SNP in all three isolates and knowing the number of passages between Z5463 and Z5463PI, we concluded that around 25 bacterial divisions occurred in the human body.Different pilin variants were found after the in vivo passage, which expressed different properties of adhesion.Furthermore the deletion of one gene involved in LOS biosynthesis (lgtB) results in Z5463BC expressing a different LOS than the L9 immunotype of Z2491.

View Article: PubMed Central - PubMed

Affiliation: INSERM U1002, Paris, France.

ABSTRACT
A scientist in our laboratory was accidentally infected while working with Z5463, a Neisseria meningitidis serogroup A strain. She developed severe symptoms (fever, meningism, purpuric lesions) that fortunately evolved with antibiotic treatment to complete recovery. Pulse-field gel electrophoresis confirmed that the isolate obtained from the blood culture (Z5463BC) was identical to Z5463, more precisely to a fourth subculture of this strain used the week before the contamination (Z5463PI). In order to get some insights into genomic modifications that can occur in vivo, we sequenced these three isolates. All the strains contained a mutated mutS allele and therefore displayed an hypermutator phenotype, consistent with the high number of mutations (SNP, Single Nucleotide Polymorphism) detected in the three strains. By comparing the number of SNP in all three isolates and knowing the number of passages between Z5463 and Z5463PI, we concluded that around 25 bacterial divisions occurred in the human body. As expected, the in vivo passage is responsible for several modifications of phase variable genes. This genomic study has been completed by transcriptomic and phenotypic studies, showing that the blood strain used a different haemoglobin-linked iron receptor (HpuA/B) than the parental strains (HmbR). Different pilin variants were found after the in vivo passage, which expressed different properties of adhesion. Furthermore the deletion of one gene involved in LOS biosynthesis (lgtB) results in Z5463BC expressing a different LOS than the L9 immunotype of Z2491. The in vivo passage, despite the small numbers of divisions, permits the selection of numerous genomic modifications that may account for the high capacity of the strain to disseminate.

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Related in: MedlinePlus

Phenotype on cells of the different pilin variants of the blood culture strain.A. Kinetic of adhesion of the different pilin variants of Z5463BC and Z5463 on hCMEC/D3 cells. An isogenic mutant of Z5463 lacking PilE was used as a negative control (ΔpilE). B. Signalling properties of the different pilin variants of Z5463BC, capable of adhesion on hCMEC/D3 cells, were assessed by immunofluorescence: in blue DAPI stain, in red actin, in green phalloïdin. All the variants recruit cell actin and phalloïdin under over 95% of the bacterial colonies forming a typical cortical plaque.
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pone-0017145-g008: Phenotype on cells of the different pilin variants of the blood culture strain.A. Kinetic of adhesion of the different pilin variants of Z5463BC and Z5463 on hCMEC/D3 cells. An isogenic mutant of Z5463 lacking PilE was used as a negative control (ΔpilE). B. Signalling properties of the different pilin variants of Z5463BC, capable of adhesion on hCMEC/D3 cells, were assessed by immunofluorescence: in blue DAPI stain, in red actin, in green phalloïdin. All the variants recruit cell actin and phalloïdin under over 95% of the bacterial colonies forming a typical cortical plaque.

Mentions: Type IV pili is an essential bacterial attribute allowing interaction of capsulated meningococci with endothelial cells. Different pilin variants allow the bacteria to express different phenotypes of bacterial-cell interaction. Among the four pilin variants directly isolated from colonies of Z5463BC, one has a non-piliated phenotype (Z5463BC2) and among the three other sequences, two were identical at the protein level. The three different derivatives were assessed for their ability to interact with cells (Fig. 8A) and to signal to cells (Fig. 8B). Both piliated isolates adhered to human brain microvessel endothelial cells (hCMEC/D3) and induced the recruitment of focal adhesion plaques with an index of recruitment superior to 95% (Fig. 8 B; data not shown). No major difference was observed between these variants except that they had varying abilities to form microcolonies onto the apical surface of the cells.


Genotypic and phenotypic modifications of Neisseria meningitidis after an accidental human passage.

Omer H, Rose G, Jolley KA, Frapy E, Zahar JR, Maiden MC, Bentley SD, Tinsley CR, Nassif X, Bille E - PLoS ONE (2011)

Phenotype on cells of the different pilin variants of the blood culture strain.A. Kinetic of adhesion of the different pilin variants of Z5463BC and Z5463 on hCMEC/D3 cells. An isogenic mutant of Z5463 lacking PilE was used as a negative control (ΔpilE). B. Signalling properties of the different pilin variants of Z5463BC, capable of adhesion on hCMEC/D3 cells, were assessed by immunofluorescence: in blue DAPI stain, in red actin, in green phalloïdin. All the variants recruit cell actin and phalloïdin under over 95% of the bacterial colonies forming a typical cortical plaque.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3046118&req=5

pone-0017145-g008: Phenotype on cells of the different pilin variants of the blood culture strain.A. Kinetic of adhesion of the different pilin variants of Z5463BC and Z5463 on hCMEC/D3 cells. An isogenic mutant of Z5463 lacking PilE was used as a negative control (ΔpilE). B. Signalling properties of the different pilin variants of Z5463BC, capable of adhesion on hCMEC/D3 cells, were assessed by immunofluorescence: in blue DAPI stain, in red actin, in green phalloïdin. All the variants recruit cell actin and phalloïdin under over 95% of the bacterial colonies forming a typical cortical plaque.
Mentions: Type IV pili is an essential bacterial attribute allowing interaction of capsulated meningococci with endothelial cells. Different pilin variants allow the bacteria to express different phenotypes of bacterial-cell interaction. Among the four pilin variants directly isolated from colonies of Z5463BC, one has a non-piliated phenotype (Z5463BC2) and among the three other sequences, two were identical at the protein level. The three different derivatives were assessed for their ability to interact with cells (Fig. 8A) and to signal to cells (Fig. 8B). Both piliated isolates adhered to human brain microvessel endothelial cells (hCMEC/D3) and induced the recruitment of focal adhesion plaques with an index of recruitment superior to 95% (Fig. 8 B; data not shown). No major difference was observed between these variants except that they had varying abilities to form microcolonies onto the apical surface of the cells.

Bottom Line: By comparing the number of SNP in all three isolates and knowing the number of passages between Z5463 and Z5463PI, we concluded that around 25 bacterial divisions occurred in the human body.Different pilin variants were found after the in vivo passage, which expressed different properties of adhesion.Furthermore the deletion of one gene involved in LOS biosynthesis (lgtB) results in Z5463BC expressing a different LOS than the L9 immunotype of Z2491.

View Article: PubMed Central - PubMed

Affiliation: INSERM U1002, Paris, France.

ABSTRACT
A scientist in our laboratory was accidentally infected while working with Z5463, a Neisseria meningitidis serogroup A strain. She developed severe symptoms (fever, meningism, purpuric lesions) that fortunately evolved with antibiotic treatment to complete recovery. Pulse-field gel electrophoresis confirmed that the isolate obtained from the blood culture (Z5463BC) was identical to Z5463, more precisely to a fourth subculture of this strain used the week before the contamination (Z5463PI). In order to get some insights into genomic modifications that can occur in vivo, we sequenced these three isolates. All the strains contained a mutated mutS allele and therefore displayed an hypermutator phenotype, consistent with the high number of mutations (SNP, Single Nucleotide Polymorphism) detected in the three strains. By comparing the number of SNP in all three isolates and knowing the number of passages between Z5463 and Z5463PI, we concluded that around 25 bacterial divisions occurred in the human body. As expected, the in vivo passage is responsible for several modifications of phase variable genes. This genomic study has been completed by transcriptomic and phenotypic studies, showing that the blood strain used a different haemoglobin-linked iron receptor (HpuA/B) than the parental strains (HmbR). Different pilin variants were found after the in vivo passage, which expressed different properties of adhesion. Furthermore the deletion of one gene involved in LOS biosynthesis (lgtB) results in Z5463BC expressing a different LOS than the L9 immunotype of Z2491. The in vivo passage, despite the small numbers of divisions, permits the selection of numerous genomic modifications that may account for the high capacity of the strain to disseminate.

Show MeSH
Related in: MedlinePlus